Identification of Proteomic Biomarkers in Proliferative Verrucous Leukoplakia through Liquid Chromatography With Tandem Mass Spectrometry

Esteban Arroyo; Mario Pérez Sayáns; Susana Belen Bravo; Camila de Oliveira Barbeiro; Mariana Paravani Palaçon; Cintia M Chamorro Petronacci; María García Vence; María Del Pilar Chantada Vázquez; Andrés Blanco Carrión; José M Suárez Peñaranda; Abel García García; Pilar Gándara Vila; Janete Días Almeida; Giovani Carlo Veríssimo da Costa; Fábio César Sousa Nogueira; Joseph Albert Medeiros Evaristo; Denise de Abreu Pereira; Mirjami Rintala; Tuula Salo; Jaana Rautava; Elena Padín Iruegas; Monica G Oliveira Alves; Túlio Morandin Ferrisse; Heitor Albergoni da Silveira; Jorge Esquiche León; Evânio Vilela Silva; Isadora Luana Flores; Andreia Bufalino

doi:10.1016/j.labinv.2023.100222

Identification of Proteomic Biomarkers in Proliferative Verrucous Leukoplakia through Liquid Chromatography With Tandem Mass Spectrometry

Lab Invest. 2023 Oct;103(10):100222. doi: 10.1016/j.labinv.2023.100222. Epub 2023 Jul 26.

Authors

Esteban Arroyo¹, Mario Pérez Sayáns², Susana Belen Bravo³, Camila de Oliveira Barbeiro¹, Mariana Paravani Palaçon¹, Cintia M Chamorro Petronacci³, María García Vence³, María Del Pilar Chantada Vázquez³, Andrés Blanco Carrión⁴, José M Suárez Peñaranda⁵, Abel García García⁴, Pilar Gándara Vila⁶, Janete Días Almeida⁷, Giovani Carlo Veríssimo da Costa⁸, Fábio César Sousa Nogueira⁹, Joseph Albert Medeiros Evaristo⁹, Denise de Abreu Pereira¹⁰, Mirjami Rintala¹¹, Tuula Salo¹², Jaana Rautava¹³, Elena Padín Iruegas¹⁴, Monica G Oliveira Alves¹⁵, Túlio Morandin Ferrisse¹⁶, Heitor Albergoni da Silveira¹, Jorge Esquiche León¹⁷, Evânio Vilela Silva¹, Isadora Luana Flores¹⁸, Andreia Bufalino¹

Affiliations

¹ Department of Diagnosis and Surgery, Araraquara, School of Dentistry, São Paulo State University (UNESP), Araraquara, São Paulo, Brazil.
² Oral Medicine, Oral Surgery and Implantology Unit (MedOralRes), Faculty of Medicine and Dentistry, University of Santiago de Compostela, Instituto de los materiales de Santiago de Compostela (iMATUS), Santiago, Spain; Instituto de Investigación Sanitaria de Santiago (IDIS) (ORALRES Group), Santiago de Compostela, Spain. Electronic address: mario.perez@usc.es.
³ Proteomic Unit, Instituto de Investigación Sanitaria de Santiago (IDIS), Santiago de Compostela, Spain.
⁴ Oral Medicine, Oral Surgery and Implantology Unit (MedOralRes), Faculty of Medicine and Dentistry, University of Santiago de Compostela, Instituto de los materiales de Santiago de Compostela (iMATUS), Santiago, Spain.
⁵ Servicio de Anatomia Patológica, Hospital Clinico Universitario de Santiago, Choupana s/n Santiago de Compostela, Spain.
⁶ Oral Medicine, Oral Surgery and Implantology Unit (MedOralRes), Faculty of Medicine and Dentistry, University of Santiago de Compostela, Instituto de los materiales de Santiago de Compostela (iMATUS), Santiago, Spain; Instituto de Investigación Sanitaria de Santiago (IDIS) (ORALRES Group), Santiago de Compostela, Spain.
⁷ Department of Bioscience and Buccal Diagnosis, São José dos Campos, Science and Technologies Institute, São Paulo State University (Unesp), São José dos Campos, São Paulo, Brazil.
⁸ Department of Basic Sciences, Nova Friburgo Health Institute, Univ. Federal Fluminense, Nova Friburgo, Rio de Janeiro, Brazil; Laboratory of Proteomics, Technological Development Support Laboratory (LADETEC), Institute of Chemistry, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
⁹ Laboratory of Proteomics, Technological Development Support Laboratory (LADETEC), Institute of Chemistry, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
¹⁰ Program on Cellular and Molecular Oncobiology, Research Coordination, National Institute of Cancer (INCA), Rio de Janeiro, Brazil.
¹¹ Department of Oral Pathology, University of Turku, Turku, Finland.
¹² Department of Oral and Maxillofacial Diseases, Clinicum, University of Helsinki, Helsinki, Finland; Department of Pathology, HUSLAB, Helsinki, Finland; Department of Cancer and Translational Research Unit, University of Oulu, Oulu, Finland; Medical Research Center Oulu, Oulu, Finland.
¹³ Department of Oral Pathology, University of Turku, Turku, Finland; Department of Oral and Maxillofacial Diseases, Clinicum, University of Helsinki, Helsinki, Finland; Department of Pathology, HUSLAB, Helsinki, Finland.
¹⁴ Human Anatomy and Embryology Area, Faculty of Physiotherapy, Department of Functional Biology and Health Sciences, Pontevedra, Spain.
¹⁵ Anhembi Morumbi University, School of Medicine, São José dos Campos, São Paulo, Brazil.
¹⁶ Department of Dental Materials and Prosthodontics, São Paulo State University (UNESP), School of Dentistry, Araraquara, Brazil.
¹⁷ Oral Pathology, Department of Stomatology, Public Oral Health, and Forensic Dentistry, Ribeirão Preto Dental School (FORP/USP), University of São Paulo, Ribeirão Preto, São Paulo, Brazil.
¹⁸ Oral Pathology Area, Conservative Dentistry Department, Dental School, Federal University of Rio Grande do Sul (UFRGS), Porto Alegre/RS, Brazil.

PMID: 37507024
DOI: 10.1016/j.labinv.2023.100222

Abstract

Proliferative verrucous leukoplakia (PVL) is an oral potentially malignant disorder associated with high risk of malignant transformation. Currently, there is no treatment available, and restrictive follow-up of patients is crucial for a better prognosis. Oral leukoplakia (OL) shares some clinical and microscopic features with PVL but exhibits different clinical manifestations and a lower rate of malignant transformation. This study aimed to investigate the proteomic profile of PVL in tissue and saliva samples to identify potential diagnostic biomarkers with therapeutic implications. Tissue and saliva samples obtained from patients with PVL were compared with those from patients with oral OL and controls. Label-free liquid chromatography with tandem mass spectrometry was employed, followed by qualitative and quantitative analyses, to identify differentially expressed proteins. Potential biomarkers were identified and further validated using immunohistochemistry. Staining intensity scan analyses were performed on tissue samples from patients with PVL, patients with OL, and controls from Brazil, Spain, and Finland. The study revealed differences in the immune system, cell cycle, DNA regulation, apoptosis pathways, and the whole proteome of PVL samples. In addition, liquid chromatography with tandem mass spectrometry analyses showed that calreticulin (CALR), receptor of activated protein C kinase 1 (RACK1), and 14-3-3 Tau-protein (YWHAQ) were highly expressed in PVL samples. Immunohistochemistry validation confirmed increased CARL expression in PVL compared with OL. Conversely, RACK1 and YWHA were highly expressed in oral potentially malignant disorder compared to the control group. Furthermore, significant differences in CALR and RACK1 expression were observed in the OL group when comparing samples with and without oral epithelial dysplasia, unlike the PVL. This research provides insights into the molecular mechanisms underlying these conditions and highlights potential targets for future diagnostic and therapeutic approaches.

Keywords: calreticulin; leukoplakia; proliferative verrucous leukoplakia; proteomic.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Biomarkers
Cell Transformation, Neoplastic / pathology
Chromatography, Liquid
Humans
Leukoplakia, Oral / diagnosis
Leukoplakia, Oral / pathology
Leukoplakia, Oral / therapy
Mouth Neoplasms* / pathology
Proteomics
Tandem Mass Spectrometry

Substances

Biomarkers